Gel electrophoresis(l15 & l16)

Question 1

Top (stacking) gel

Answer 1

%5 acrylamide, pH 6.8 with tris-Cl

Question 2

Bottom(running) gel

Answer 2

12% acrylamide, pH 8.8 w/ tris-cl

Question 3

Upper buffer

Answer 3

pH 8.3 w/ glyce (pI =6)

Question 4

Rf

Answer 4

how fast your protein dye goes compared to something fast(dye)
- removes problems of absolute distances when 2 gels are run at different times/voltages

Question 5

Rf vs molecular mass

Answer 5

gives a protein size molecular curve

Question 6

acrylamide mobility and size of proteins(questionize this)

Answer 6

mobility increases exponentially as size decreases

Question 7

SDS-page allows for

Answer 7

seperation of proteins based on a single criteria(size)

Question 8

urea gels

Answer 8

• proteins denature but retain their native charge
• seperate proteins by size and charge
• run at specific pH to control protein charges

Question 9

Acid urea gel

Answer 9

5% acetic acid
protonates some R groups(overall positive charge)
high acrylamide
slower than SDS and backwards

Question 10

native gels

Answer 10

mobility depends on size charge and conformation
charge depends on pH
substantially slower than SDS-page

Question 11

Native gel: sickle cell anemia

Answer 11

glutamic acid(-) -> valine(non-polar)

• charge differences show up nicely
• tetramer not broken

Question 12

2-dimentional protein electrophoresis

Answer 12

seperated based on 2 factors in 2 dimensions

Question 13

ampholytes buffereing at their pKa allows them to

Answer 13

maintain a pH gradient without moving during electrophoresis

Question 14

proteins in isoelectric focusing gels

Answer 14

seperate by the average pI of all ionizable groups

Question 15

isoelectric focusing strips

Answer 15

treated to denature proteins and coated with sds